This proposal describes a 5-year training program for the candidate, Catherine Hagan, D.V.M., to become an independent investigator in comparative experimental neuropathology. She is pursuing a Ph.D. in the Molecular and Cellular Biology graduate program at the University of Washington, Seattle; her dissertation work focuses on mechanisms of extracellular serotonin regulation in the brain. With this SERCA, Dr. Hagan will build on her foundation in brain serotonin biology to investigate serotonergic immunomodulation of microglial-mediated neuroinflammation with co-mentor Dr. John Neumaier, M.D., Ph.D., co-mentor Dr. Tom Montine, M.D., Ph.D., and collaborator Dr. Thomas Moller, Ph.D. Their combined expertise in serotonin pharmacology, neuropathology, and microglial biology is a unique feature of the rich training environment at the University of Washington. Career development activities will include specialized courses, such as the Molecular Neuropathology course at Cold Spring Harbor, and regular didactic activities such as a journal clubs focused on microglial biology, inflammation, and neuroscience. Microglia are the immune system of the brain and are critically involved in responses to injury and various diseases. These responses can be either beneficial or detrimental. Conditions leading to these dichotomous responses are poorly understood. The long term goal of the proposed research is to understand how serotonin and serotonin receptor mediated signaling between neurons and microglia contribute to pathogenesis of neuroinflammatory diseases. Aim 1 focuses on how the serotonin signal to microglia may be disrupted by inflammation. Rotating disk voltammetry will be used to study whether stimulating cerebral innate immunity in vivo alters neuronal serotonin transporter kinetics. This may be an underlying mechanism for how changes in serotonin lead to changes in microglial function. Aim 2 explores the possibility that microglia have an important functional capacity to take up serotonin and the mechanisms by which occurs. Aim 3 focuses on microglial serotonin receptor expression and function. Results will be used to test the hypothesis that impaired serotonergic neurotransmission and signaling potentiates microglial-mediated neurotoxicity. Immunomodulation of microglia has the potential to improve clinical outcomes in a variety of disorders, including neuroinflammatory disorders, neurodegenerative diseases, and chronic pain disorders. Accomplishing the aims of the proposed studies will determine if targeting serotonin signaling between neurons and microglia offers a new therapeutic approach to neuroinflammation through modulation of microglial responses to injury. PERFORMANCE SITE(S) (organization, city, state) The University of Washington, Seattle, WA 1. Harborview Medical Center Campus, Seattle, WA A. Neumaier Laboratory at Harborview Research and Training Building, Department of Psychiatry and Behavioral Sciences, School of Medicine, University of Washington, Seattle, WA B. Montine Laboratory at Harborview Research and Training Building, Department of Pathology, School of Medicine, University of Washington, Seattle, WA 2. Department of Comparative Medicine, School of Medicine, University of Washington, Seattle, WA PHS 398 (Rev. 04/06) Page 2 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): Hagan, Catherine, Elizabeth KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator(s). List all other key personnel in alphabetical order, last name first. Name . eRA Commons User Name Organization Role on Project Hagan, Catherine E. cehagan University of Washington Principal Investigator Liggitt, H. Denny dliggitt University of Washington Co-mentor Montine, Thomas J. tmontine University of Washington Mentor Neumaier, John F. neumaier University of Washington Mentor OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Moeller, Thomas University of Washington Collaborator Schenk, Jim O. Washington State University Collaborator Human Embryonic Stem Cells [x] No Q Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the followinglist: http://stemcells.nih.qov/reaistrv/index.asp. Use continuation pages as needed. If a specific line cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line PHS 398 (Rev. 04/06) Page 3 Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. CDA TOC Substitute Page candidate (Last, first, middle): Hagan, Catherine, Elizabeth Use this substitute page for the Table of Contents of Research Career Development Awards. Type the name of the candidate at the top of each printed page and each continuation page. RESEARCH CAREER DEVELOPMENT AWARD TABLE OF CONTENTS (Substitute Page) Page Numbers Letters of Reference* (attach unopened references to the Face Page) Section I: Basic Administrative Data Face Page (Form Page 1) 1 Description, Performance Sites, Key Personnel, Other Significant Contributors, and Human Embryonic Stem Cells (Form Page 2) 2-3 Table of Contents (this CDA Substitute Form Page 3) 4 Budget for Entire Proposed Period of Support (Form Page 5) 6-8 Biographical Sketches (Candidate, Sponsors],' Key Personnel and Other Significant Contributors' -Biographical Sketch Format page) (Not to exceed four pages) 9-29 Other Support Pages (not for the candidate) 30-32 Resources (Resources Format page) 33-37 Section II: Specialized Information Introduction to Revised/Resubmission Application* (Not to exceed 3 pages) 38-40 1. The Candidate A. Candidate's Background rry f 41 B.CareerGoalsandObjectives:ScientificBiography r...(ItemsA-Dincludedin25pagelimit) I 42-45 C. Career Development/Training Activities during Award Period pound. :.... | 45-49 D. Training in theResponsible Conduct ofResearch J ^ 49 2. Statements by Sponsor, Co-Sponsor(s),* Consultant(s),* and Contributor(s)' 50-64 3. Environment and Institutional Commitment to Candidate A. Description of Institutional Environment 65-67 B. Institutional Commitment to Candidate's Research Career Development 68 4. Research Plan A. Specific Aims T!l C 69 B. Background and Significance .L^ (Items A-D included in 25 page limit) J 70-71 C. Preliminary Studies/Progress Report I I 72-76 D. Research Design and Methods -J. V. 77-84 E. Human Subjects Research n/a Targeted/Planned Enrollment Table (for new and continuing clinical research studies) n/a F. Vertebrate Animals 85 G. Select Agent Research n/a H. Literature Cited...! 86-90 I. Consortium/Contractual Arrangements* n/a J. Resource Sharing n/a Checklist 91 Appendix (Five collated sets. No page numbering necessary.) | | Check if Appendix is included Number of publications and manuscripts accepted for publication (not to exceed 5) List of Key Items: Note: Font and margin requirements must conform to limits provided in the Specific Instructions. 'Include these items only when applicable. CITIZENSHIP E U.S. citizen or O Permanent resident of U.S. (If a permanent resident of the U.S., a d Non-citizen with temporary visa non-citizen national notarized statement must be provided by the time of award.) (Applicable for only the K99 program) PHS 398 (Rev. 04/06) Page 4 CDA Substitute Form Page 3